I want to site-directed mutation on genome other than plasmid DNA. Anyone has protocol like this. I donnot want to introduce neo etc antibiotic maker. I need the clear genetic background which the mutation differ to the wild type with only the nucleotide.
any help is wellcome!!
Thank a lot.
well first you have to figure out what your target sequence is (the sequence of the chromatin you want to mutate). Then make an artificial oligo that has the mutation, and introduce it into the genome by recombination insertion.